Richardson, Thomas
(2017)
Fundamental Study of Alginate-Based 3D Platforms for the Propagation and Pancreatic Differentiation of Human Embryonic Stem Cells.
Doctoral Dissertation, University of Pittsburgh.
(Unpublished)
Abstract
Type 1 diabetes (T1D) is an autoimmune disease affecting millions of people worldwide, wherein the insulin producing pancreatic islets are destroyed. Shortage of donor cells, combined with immune rejection, limits transplantation as a viable therapy. However, human pluripotent stem cells (hPSCs) have the capacity to become any cell type, and the potential to provide an unlimited supply of hPSC-derived insulin producing cells. Additionally, immune rejection is still a hurdle which must be overcome before hESC-based therapy for T1D can be brought to fruition. It is therefore necessary to develop suitable culture and differentiation strategies which employ cell encapsulation with an appropriate material which can protect the new insulin producing cells from the body’s immune system and is supportive of hPSC biomanufacturing.
Alginate encapsulation has been previously used for providing the needed immune protection of donor insulin-producing cells for transplantation. Therefore, in this work we first showed that alginate encapsulation supports efficient differentiation of hPSCs to insulin producing cells, and also significantly enhanced maturation of hPSCs compared to 2D controls. Previous studies for islet encapsulation have also shown that alginate hydrogel composition significantly affects the capacity for immune isolation. However, such capsule compositions could modify the differentiation of the encapsulated cells. Thus, we next evaluated the effect of capsule composition on material properties and thus on pancreatic differentiation of encapsulated hESCs. Our results clearly showed that even in the presence of chemical differentiation factors,
Fundamental Study of Alginate-Based 3D Platforms for the Propagation and Pancreatic Differentiation of Human Embryonic Stem Cells
Thomas Richardson, PhD
University of Pittsburgh, 2017
v
substrate stiffness greatly affects the efficiency of pancreatic differentiation. Taking this a step further, we developed a high throughput 3D alginate array platform that allowed for multivariate perturbations of insoluble differentiation cues, namely alginate stiffness and cell-cell contact, during pancreatic differentiation. Our results accordingly indicated that while stiffness did influence proliferation and pancreatic differentiation, the effect of cell-cell contact was more significant.
Finally, we developed an e-cadherin mimicking peptide-conjugated alginate substrate which mimics cell-cell contact, to meet the biomanufacturing needs of hPSCs. The designed biomimetic substrate supported single cell survival and propagation, as well as maintained hPSC pluripotency and differentiation potency. Overall, the findings from this dissertation represent a significant advancement in strategies supporting the propagation and pancreatic differentiation for biomanufacturing of hPSCs as an effective cell therapy treatment for T1D.
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Details
| Item Type: |
University of Pittsburgh ETD
|
| Status: |
Unpublished |
| Creators/Authors: |
|
| ETD Committee: |
|
| Date: |
26 September 2017 |
| Date Type: |
Publication |
| Defense Date: |
20 June 2017 |
| Approval Date: |
26 September 2017 |
| Submission Date: |
11 July 2017 |
| Access Restriction: |
1 year -- Restrict access to University of Pittsburgh for a period of 1 year. |
| Number of Pages: |
202 |
| Institution: |
University of Pittsburgh |
| Schools and Programs: |
Swanson School of Engineering > Chemical and Petroleum Engineering |
| Degree: |
PhD - Doctor of Philosophy |
| Thesis Type: |
Doctoral Dissertation |
| Refereed: |
Yes |
| Uncontrolled Keywords: |
Alginate, substrate stiffness, encapsulation, hESC, stem cells, diabetes, insulin |
| Date Deposited: |
26 Sep 2018 05:00 |
| Last Modified: |
26 Sep 2018 05:15 |
| URI: |
http://d-scholarship.pitt.edu/id/eprint/32740 |
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